Decidualization Potency and Epigenetic Changes in Human Endometrial Origin Stem Cells During Propagation

Abstract

Human endometrium derived mesenchymal stem cells (hEndSCs) offer a great promise for regenerative medicine and reproductive system disorders treatment methods based on cell therapy due to their broad differentiation potential and highly efficient proliferation. In our study, we investigated the characteristics of hEndSCs that were isolated from two sources: endometrium and menstrual blood, which both contain endometrial origin stem cells. Changes in gene and protein expression levels during long-term cultivation and decidualization potential were examined in endometrial stem cells (EndSCs) and menstrual blood stem cells (MenSCs). The decidualization process was induced on early and late passages of hEndSCs using dibutyryl cyclic-AMP (db-cAMP) and medroxyprogesterone acetate (MPA) agents. We demonstrated that after long-term cultivation of hEndSCs the expression of typical mesenchymal stromal cell surface markers such as CD44, CD73, CD90, CD105 and perivascular marker CD146 remains at a similar level throughout long-term cultivation. Additionally, hematopoietic and endothelial markers CD34, CD45 were also tested, they were negative in all cases. Analyzed stem cells gene markers, such as OCT4, SOX2, NANOG, KLF4, showed similar expression in all passages of hEndSCs. RT-qPCR results demonstrated that the expression of cell cycle control associated genes - CDK2, CCNA2, CCNE2, p21, p53 and Rb, among all groups was very similar. Expression of genes associated with senescence (ATM, JUND, TOP2A, MYC) was maintained at a similar level throughout passaging. In addition, Western blot analysis was used to assess changes in proteins’ levels associated to epigenetics (EZH2, SUZ12, H3K27me3) and cell cycle control (cyclinE1, p53) during long-term cultivation. The levels of proteins associated with epigenetic changes were fluctuated slightly depending on the patient. Also, we demonstrated that in all induced hEndSCs the expression of decidualization markers Prolactin (PRL), IGFBP1 and WNT4 was upregulated. In conclusion, we demonstrated successful decidualization of stem cells derived from two reproductive system resources: endometrium and menstrual blood by using db-cAMP and MPA regardless of the length of the stem cell passaging. According these findings, we suppose that endometrium derived stem cells and menstrual blood derived stem cells could have a potency not only for endometrium tissue regeneration, but could also become a successful therapy for reproductive system disorders, including infertility or recurrent pregnancy loss.

The Enzyme 15-Hydroxyprostaglandin Dehydrogenase Inhibits a Shift to the Mesenchymal Pattern of Trophoblasts and Decidual Stromal Cells Accompanied by Prostaglandin Transporter in Preeclampsia

“…Human decidual stromal cells (hESCs) were induced to decidualize (decidualized hESC: dhESC) according to the above methods [ 14 ]. Western blots revealed that prolactin (PRL) and insulin-like growth factor binding protein 1 (IGFBP1), the traditional markers of decidualization, increased ( Figure 1 E), indicating that in vitro hESC decidualization was successful; moreover, CDH1 increased, and vimentin decreased during hESC decidualization in vitro ( Figure 1 E,G).…”

“…hESCs were cultured in a 5% humidified carbon dioxide atmosphere at 37 °C in phenol red-free Dulbecco’s modified Eagle’s medium (DMEM)/F-12 (Meilunbio, Dalian, China) with 10% foetal bovine serum, 50 mg/mL streptomycin, and 50 U/mL penicillin. One micromolar medroxyprogesterone-17-acetate (MPA) (HY-B0469, MedChemExpress, NJ, USA) and 0.5 mM N6,20-O-dibutyryladenosine cAMP sodium salt (db-cAMP) (HY-B0764, MedChemExpress, Monmouth Junction, NJ, USA) were added to the culture for 6 days to induce hESC decidualization in vitro [ 14 ].…”

The role of human endometrial stem cells application in endometrial-factor induced infertility

“…These ndings could be explained by the activity of hEnSCs. They are characterized by the broad differentiation potential, highly e cient self-renewal and remarkable immunomodulatory capacity to migrate to the injured reproductive tissues caused by chemokines, before they differentiate and integrate somatic cells with non-tumorigenic properties [22,57,58]. It creates conditions for slowing down the progression of brosis and (or) replacing damaged tissues [58-60].…”

Efficient bone regeneration of BMP9-stimulated human periodontal ligament stem cells (hPDLSCs) in decellularized bone matrix (DBM) constructs to model maxillofacial intrabony defect repair

“…The expression of stem cell surface markers of hDMSCs was characterized by means of flow cytometry. The fluorescence-labeled antibodies were CD90-FITC (Sino Biological, China), CD29-FITC (Sino Biological, China) and CD45-FITC (Sino Biological, China) as previously described [31][32][33].…”